Epifluorescence Microscopy

The platform has two widefield microscopes for the acquisition of material fixed in fluorescence, a widefield microscope equipped with a colour camera, a routine fluorescence microscope for the observation of living cells and an analysis computer equipped with Fiji and Icy software. This equipment is reserved for the internal use of UMR7216 members.

Artistic image of jellyfish, referring to the bioluminescent jellyfish from which GFP (Green Fluorescent Protein) is extracted that has revolutionised the world of imaging.

© Image par Carol-Ann Bussières de Pixabay

Leica DMI-6000B "Oldelaf"

Inverted epifluorescence microscope
X, Y and Z piloted
Filters and objectives piloted

Camera:
Camera CCD -30ºC regulated
CoolSNAP HQ2 14bits

Image size: 1392×1040 pixels
Pixel size: 6.45 x 6.45μm

Illumination:
Visible light (halogen)
CoolLED pE-300 White Series Fluorescent Lamp (LED, >25000h)

Acquisition software:
Metamorph with Multi-Dimensional Acquisition module

Objectives:

Obj Mag. N.A Quality Transmission Contrast Imm. Pixel size (binning 1) Max resolution at 488nm
10X 0,25 N PLAN PH1 Air 0,645 μm 1191 nm
20X 0,35 N PLAN PH1 Air 0,3225 μm 850 nm
40X 0,6 HCX PL Fluotar PH2 Air 0,161 μm 496 nm
40X 1,25-0,75 HC PL APO CS – Oil 0,161 μm 229 nm
63X 1,40 – 0.60 HCX PL APO – Oil 0,102 μm 213 nm
100X 1,40 HCL PL APO CS PH3 Oil 0,0645 μm 213 nm

Filters :

Filter name Colour Excitation Filter BP Dichroic Mirror Suppression Filter BP
A4 Blue 360/40 400 470/40
GFP Green 470/40 500 525/50
Y3 Cy3 (orange) 545/40 565 610/75
TX2 Red 560/40 595 645/75
Y5 Cy5 (Far red) 620/60 660 700/75
Analyzer cube – – – Analyzer

 

Leica DMI-6000B "Lenny"

Inverted epifluorescence microscope
Controlled in X, Y and Z
Controlled filters and lenses

Camera:
Camera CCD -12ºC regulated
Teledyne Photometrics RETIGA R6

Image size: 2688×2200 pixels
Pixel size: 4.54 x 4.54μm

Illumination:
Visible light (halogen)
CoolLED pE-300 White Series Fluorescent Lamp (LED, >25000h)

Acquisition software:
Metamorph with Multi-Dimensional Acquisition module

Objectives:

Mag. N.A Quality Trans Contrast Imm. Pixel size (binning 1) Max resolution at 488 nm
2,5X 0,07 HC FL PLAN – Air 1.816 μm 4,2 µm
20X 0,7 HCL PL APO CORR – Oil/Gly 0,227 μm 425 nm
40X 1,30 HC PL APO – Oil 0,1135 μm 229 nm
63X 1,40 HCX PL APO CS PH3 Oil 0,072 μm 212 nm
100X 1,40-0,7 HCX PL APO – Oil 0,0454 μm 212 nm

Filters :

Name Colour Excitation Filter BP Dichroic Mirror Suppression Filter BP
XF131 Blue 387/28 410 450/65
QMAX green Green 450-490 500 510-560
N3 Cy3 (orange) 546/12 565 600/40
TX2 Red 560/40 595 645/75
Far Red Cy5 620-60 660 700-75
Analyzer cube – – – Analyzer
Leica DM IL LED (Live)

Inverted epifluorescence microscope

Camera:
Camera CCD -60ºC regulated
Hamamatsu digital camera C4742-98-24ERG 12 or 14 bits

Image size: 1344×1024
Pixel size: 6.45 x 6.45μm

Illumination:
Visible light (halogen)
Leica fluorescent lamp (Mercury Halides – 3000h max)

Objectives:

Mag. N.A Quality Transmission
Contrast		 Imm. Pixel size
(binning 1)		 Max resolution at 488 nm
10X 0,25 N PLAN PH1 Air 0,645 μm 1,2 µm
20X 0,35 N PLAN PH1 Air 0,322 μm 850 nm
40X 0,55 CORR PH2 Air 0,161 μm 541 nm

Filtres :

Name Colour Excitation Filter BP Dichroic Mirror Suppression Filter BP
B/G/R Blue/Green/Red 420/30 ; 495 /15 ; 570/20 415 ; 510 ; 590 465/20 ; 530 /30 ; 640/40
GFP ET Green      
TX ET Red      

 

 

Leica DMRA2 (Color)

Upright epifluorescence microscope
Controlled in X, Y and Z

Camera:
CCD digital color camera 5Mpixels Leica DFC 450C cooled (Δ -20ºC compared to ambient)

Image size: 2560×1920
Pixel size: 3.4 x 3.4μm

Illumination:
Visible light (halogen)
Fluorescent lamp not installed

Objectives:

Mag. N.A Quality Transmission
Contrast		 Imm. Pixel size
(binning 1)		 Max resolution at 488 nm
10X 0,30 HC PL Fluotar – Air 0,34 μm 992 nm
20X 0,50 HCX PL Fluotar – Air 0,17 μm 595 nm
40X 0,75 HCX PL Fluotar – Air 0,085 μm 397 nm
100X 1,40-0,7 HCX PL APO CS – Oil 0,034 μm 212 nm
Workstation "Angus"

A workstation for image analysis is available. This workstation is equipped with the free software FIJI, IMAGEJ, ICY.

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